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LakePharma

Belmont, California, US

LakePharma is a leading contract research organization (CRO) specializing in high-quality antibody and protein engineering, expression and purification, cell line development, and DNA cloning and production. LakePharma's services are fully integrated, creating a streamlined solution for all protein engineering needs.

• Founded in 2009 - Nearly 100 employees, 25% with Ph.D.s
• Located in both San Francisco Bay Area and Greater Boston Area - all work is completed in U.S.
• Key technologies developed in-house for efficient and high quality services
• Quality control in place
• Dedicated client services team to ensure client satisfaction
• Secure, cloud-based online Datasystems
• GLP services available upon request

LakePharma is a leading contract research organization (CRO) specializing in high-quality antibody and protein engineering, expression and purification, cell line development, and DNA cloning and production. LakePharma's services are fully integrated, creating a streamlined solution for all protein engineering needs.

• Founded in 2009 - Nearly 100 employees, 25% with Ph.D.s
• Located in both San Francisco Bay Area and Greater Boston Area - all work is completed in U.S.
• Key technologies developed in-house for efficient and high quality services
• Quality control in place
• Dedicated client services team to ensure client satisfaction
• Secure, cloud-based online Datasystems
• GLP services available upon request

Standard sample shipment requirements:
- All packages must be shipped overnight with the appropriate packaging and labeling.
- All samples must be shipped with a description and an MSDS safety sheet if applicable.

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Plasmid DNA Preparation

Starting at $150.00 per prep

LakePharma produces endotoxin-free plasmid DNA at multiple scales. We can turn around a prep from a bacterial pellet in 24 hours (see below for details). The endotoxin levels in all delivered endotoxin-free samples are below 0.01 EU per microgram of DNA. Final products are verified with either restriction enzyme digest or sequence... Show more »

LakePharma produces endotoxin-free plasmid DNA at multiple scales. We can turn around a prep from a bacterial pellet in 24 hours (see below for details). The endotoxin levels in all delivered endotoxin-free samples are below 0.01 EU per microgram of DNA. Final products are verified with either restriction enzyme digest or sequence analysis.

Endotoxin-free DNA is critical for sensitive cell culture and in vivo experiments. LakePharma produces virtually endotoxin-free plasmid DNA at multiple scales. The endotoxin levels in all delivered endotoxin-free samples are below 100 EU per milligram of DNA. Final products are verified with either restriction enzyme digest or sequence analysis.

Monoclonal antibodies to clinically important target proteins have been developed and used as therapies to a wide variety of diseases. Monoclonal antibodies are usually produced in non-human species, most often in mice. However, to become successful biotherapeutics, these non-human antibodies have to overcome several significant... Show more »

Monoclonal antibodies to clinically important target proteins have been developed and used as therapies to a wide variety of diseases. Monoclonal antibodies are usually produced in non-human species, most often in mice. However, to become successful biotherapeutics, these non-human antibodies have to overcome several significant drawbacks. First, non-humanized antibodies can elicit an immunogenic response when administered to patients, as the human immune system can recognize the non-humanized antibodies as foreign. Creation of chimeric antibodies (non-human variable regions with human constant regions) can partially alleviate these problems. A much more effective strategy is humanization of the antibody. This process involves transferring the antibody CDRs into a proper human antibody framework, while retaining its original affinity and specificity. LakePharma has developed and validated an efficient platform for antibody humanization that combines the advantages of both rational and empirical approaches.

Antibodies can be developed through classic hybridoma technology; as an alternative to hybridoma development, LakePharma offers construction and screening of scFv libraries using phage display. Once a hybridoma that produces antibodies with the desired properties is identified, the heavy and light chain variable regions can be cloned, which establishes the identity of the antibodies and enables the expression and production of full-length recombinant antibodies.

In addition to hybridomas from multiple species, clonal B cells, including single cells, can be used as the starting point for molecular cloning. LakePharma can also provide de novo sequencing of an antibody by mass spectrometry when the starting material is a pure monoclonal antibody and the source of DNA is not available.

LakePharma offers a broad range of antibody engineering and characterization assays for lead identification and optimization. We have extensive experience in reconfiguring antibodies into a variety of formats and rapid production at various scales. Additionally, we routinely perform antibody humanization, affinity maturation and... Show more »

LakePharma offers a broad range of antibody engineering and characterization assays for lead identification and optimization. We have extensive experience in reconfiguring antibodies into a variety of formats and rapid production at various scales. Additionally, we routinely perform antibody humanization, affinity maturation and optimization, creating antibodies for numerous applications, including therapeutics.

Binding confirmation and affinity measurement of the antibodies produced can be performed using an assortment of methods, including Octet (biolayer interferometry BLI), ELISA and flow cytometry.

In conjunction with antibody generation, protein analytics, and cell line development services, the LakePharma antibody engineering and production platform provides end-to-end service for antibody discovery and development.

LakePharma has extensive experience with constructing and producing antibodies in our mammalian platforms. Antibody variable region genes (VH and VL) from a given antibody can be reformatted to encode a different constant region and other antibody-like molecules including Fab, scFv, and bispecific molecules. Multiple species and... Show more »

LakePharma has extensive experience with constructing and producing antibodies in our mammalian platforms. Antibody variable region genes (VH and VL) from a given antibody can be reformatted to encode a different constant region and other antibody-like molecules including Fab, scFv, and bispecific molecules. Multiple species and isotypes are available, with the option of synthesizing the constant region of choice. These expression constructs can be expressed in HEK293 and CHO cells followed by protein purification, and rapidly produced at scales ranging from milligram to gram quantities. We also offer high throughput 96-block antibody productions to fit your antibody screening needs.

Gene synthesis is the process of assembling a coding sequence from individual nucleotides or short oligos. Using our Chain Blue Lightning™ methodologies, we are able to quickly and efficiently synthesize DNA molecules of varying sizes. Gene synthesis has emerged as a more rapid and dynamic alternative tool for genetic manipulation... Show more »

Gene synthesis is the process of assembling a coding sequence from individual nucleotides or short oligos. Using our Chain Blue Lightning™ methodologies, we are able to quickly and efficiently synthesize DNA molecules of varying sizes. Gene synthesis has emerged as a more rapid and dynamic alternative tool for genetic manipulation than PCR cloning and is able to provide researchers with benefits that include:

Improved expression: the gene sequence can be optimized for expression in specific organisms.
No restrictions: Gene can be designed for cloning into any expression vector.
Save time: Make multiple mutations or variants faster and easier than using traditional PCR cloning methods.
Save money: Gene synthesis is often more economical than PCR cloning.

Our team offers a diverse variety of biochemical and cell based assay development and screening solutions. We have the knowledge and flexibility to support your assay needs. Our dedicated team of experienced scientists will work with you to transfer and validate an existing assay, or develop a custom assay to your specifications.... Show more »

Our team offers a diverse variety of biochemical and cell based assay development and screening solutions. We have the knowledge and flexibility to support your assay needs. Our dedicated team of experienced scientists will work with you to transfer and validate an existing assay, or develop a custom assay to your specifications. In our state of the art facility, we can perform radiometric, fluorescence, image-based, and other screening formats using your compounds, or our proprietary focused library. In addition, our project leaders will guide you through the entire process, notifying you about developments every step of the way.

From one plate studies to recurrent compound testing and 100’s of plates, our team can deliver the high quality data that you want with the fast turnaround times you demand.

Gene synthesis is the process of assembling a coding sequence from individual nucleotides or short oligos. Using our Chain Blue Lightning™ methodologies, we are able to quickly and efficiently synthesize DNA molecules of varying sizes. Gene synthesis has emerged as a more rapid and dynamic alternative tool for genetic manipulation... Show more »

Gene synthesis is the process of assembling a coding sequence from individual nucleotides or short oligos. Using our Chain Blue Lightning™ methodologies, we are able to quickly and efficiently synthesize DNA molecules of varying sizes. Gene synthesis has emerged as a more rapid and dynamic alternative tool for genetic manipulation than PCR cloning and is able to provide researchers with benefits that include:

Improved expression: the gene sequence can be optimized for expression in specific organisms.

No restrictions: Gene can be designed for cloning into any expression vector.

Save time: Make multiple mutations or variants faster and easier than using traditional PCR cloning methods.

The Protein Sciences’ team employs a variety of chromatographic methodologies on industry standard platforms to purify recombinant and endogenous proteins from many sources, including tissue/organs, cell paste, and conditioned media.

LakePharma’s expert scientists routinely replicate established protocols or conduct process... Show more »

The Protein Sciences’ team employs a variety of chromatographic methodologies on industry standard platforms to purify recombinant and endogenous proteins from many sources, including tissue/organs, cell paste, and conditioned media.

LakePharma’s expert scientists routinely replicate established protocols or conduct process development in cases where protocols do not exist. Wherever applicable, our team is proactive in recommending improvements to existing protocols.

Quantitative Real Time PCR (qPCR) is an important research tool for quantifying gene expression or DNA copy number. We offers rapid, high quality qPCR analysis using our state of the art equipment. Our full service qPCR offering includes RNA and cDNA preparation, primer design, experimental design, qPCR experiment execution and... Show more »

Quantitative Real Time PCR (qPCR) is an important research tool for quantifying gene expression or DNA copy number. We offers rapid, high quality qPCR analysis using our state of the art equipment. Our full service qPCR offering includes RNA and cDNA preparation, primer design, experimental design, qPCR experiment execution and data analysis, incluing normalization to internal controls. LakePharma utilizes both general fluorescent dyes (ex. SYBR Green) and gene specific probes.

Site-Directed Mutagenesis is an important research tool for protein engineering and structure/function studies. Our mutagenesis service can be conducted using Chain Blue Lightning™ Gene Synthesis or PCR Nucleotide Substitutionmediated mutagenesis and allows rational design of protein variants using variety methodologies, including:

Scanning Mutagenesis

Deletions and Insertions

Site-Directed Mutagenesis is an important research tool for protein engineering and structure/function studies. Our mutagenesis service can be conducted using Chain Blue Lightning™ Gene Synthesis or PCR Nucleotide Substitutionmediated mutagenesis and allows rational design of protein variants using variety methodologies, including:

Expression of recombinant proteins in E. coli is often the fastest and most cost-effective solution for recombinant protein production. We leverage many years of experience in bacterial expression services to deliver the highest quality recombinant protein.

Expression of recombinant proteins in E. coli is often the fastest and most cost-effective solution for recombinant protein production. We leverage many years of experience in bacterial expression services to deliver the highest quality recombinant protein.

Our Bacterial Expression services include:

Strain creation

Expression optimization (temperature and induction conditions)

Flask and Fermentor production

batch, fed-batch and continuous culture

10-50L in 5-15 business days

Process Development

Bacterial protein production of His-tag protein - Starting at $2,500.00 per sample

Yeast systems allow efficient production of secreted and intracellular targets. In addition, yeast provides the advantage of post-translational modifications, which may be beneficial for some recombinant proteins.
The Yeast Expression services include:

Yeast systems allow efficient production of secreted and intracellular targets. In addition, yeast provides the advantage of post-translational modifications, which may be beneficial for some recombinant proteins.
The Yeast Expression services include:

With the increasing popularity of large-scale mammalian transient protein expression, there has been a concomitant demand for bulk DNA production. The plasmid TeraPrep™ service fulfills this demand with a large-scale chromatographic DNA purification platform developed by our molecular biology and protein science teams. It provides... Show more »

With the increasing popularity of large-scale mammalian transient protein expression, there has been a concomitant demand for bulk DNA production. The plasmid TeraPrep™ service fulfills this demand with a large-scale chromatographic DNA purification platform developed by our molecular biology and protein science teams. It provides high-purity, endotoxin-free DNA with unmatched turnaround times of one – two weeks in milligram to gram quantities.
The TeraPrep™ Service provides:

Reverse Transcriptase PCR (RT-PCR) is a technique where mRNAs are reverse transcribed into cDNA using the enzyme reverse transcriptase. The resulting cDNA can be used for a variety of applications including qPCR, sequence validation or expression. cDNA can be amplified using standard PCR and cloned directly into cloning or expression vectors.

Reverse Transcriptase PCR (RT-PCR) is a technique where mRNAs are reverse transcribed into cDNA using the enzyme reverse transcriptase. The resulting cDNA can be used for a variety of applications including qPCR, sequence validation or expression. cDNA can be amplified using standard PCR and cloned directly into cloning or expression vectors.

Additionally, we offer complete antibody production services, including cloning of heavy and light chain variable regions or full-length antibodies, co-expression, purification and validation of antigen binding by ELISA.

Although the amount available genomic data has grown exponentially over the last few years, there remain many organisms of interest without reliable genomic data. LakePharma offers bioinformatic and de novo cDNA services to identify and clone gene orthologs. Our experienced team provides comprehensive cDNA cloning services... Show more »

Although the amount available genomic data has grown exponentially over the last few years, there remain many organisms of interest without reliable genomic data. LakePharma offers bioinformatic and de novo cDNA services to identify and clone gene orthologs. Our experienced team provides comprehensive cDNA cloning services enhancing your ability to produce a variety of proteins.

Save time: Clone your novel gene of interest into an expression vector and advancing it directly into expression testing without delay.

For downstream applications of recombinant proteins, yield and purity are not the only metrics by which proteins preparations are judged. Frequently, proteins must also have low endotoxin levels because these molecules interfere with cellular assays and animal studies. To address this important issue, the Protein Sciences team... Show more »

For downstream applications of recombinant proteins, yield and purity are not the only metrics by which proteins preparations are judged. Frequently, proteins must also have low endotoxin levels because these molecules interfere with cellular assays and animal studies. To address this important issue, the Protein Sciences team has developed the DeTox™ platform, whereby recombinant proteins are subjected to a rigorous process of endotoxin minimization, testing, and removal (if necessary). This endotoxin-control system delivers recombinant protein without the hassles of contaminating endotoxin, irrespective of the expression system used.

The LakePharma team uses a membrane-based Tangential Flow Filtration (TFF) system to clarify, concentrate and enrich proteins, which reduces downstream processing times. Volumes as large as 300L can be processed in a matter of a few hours at a several molecular weight cutoffs, allowing the separation of an extensive size range of... Show more »

The LakePharma team uses a membrane-based Tangential Flow Filtration (TFF) system to clarify, concentrate and enrich proteins, which reduces downstream processing times. Volumes as large as 300L can be processed in a matter of a few hours at a several molecular weight cutoffs, allowing the separation of an extensive size range of macromolecules, including viruses.

Fluorescence- and luminescence-based assays have become increasingly popular in recent years with a wide diversity of options available for different needs. Blue Sky offers custom assay development and screening services using Fluorescence Polarization (FP), Fluorescence Resonance Energy Transfer (FRET), Time-Resolved Fluorescence... Show more »

Fluorescence- and luminescence-based assays have become increasingly popular in recent years with a wide diversity of options available for different needs. Blue Sky offers custom assay development and screening services using Fluorescence Polarization (FP), Fluorescence Resonance Energy Transfer (FRET), Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET), Homogeneous Time Resolved Fluorescence (HTRF), and luminescence-based assays.

Antibody-based assays are popular in drug discovery and biomedical research to detect specific components in complex systems. LakePharma offers custom solutions to suit your needs for development and execution of these assays. We have the capability and expertise to help you quantify yields from your expression and purification... Show more »

Antibody-based assays are popular in drug discovery and biomedical research to detect specific components in complex systems. LakePharma offers custom solutions to suit your needs for development and execution of these assays. We have the capability and expertise to help you quantify yields from your expression and purification endeavors, assist in clonal selection, and even compare post-translational modification states of proteins. Our scientists also have the skills to free your time by developing and optimizing custom ELISA and immunoassay solutions to assist in your unique workflow.

Radiometric assays are the “gold standard” for many laboratories but establishing and maintaining these capabilities can be a burden for clients with limited resources. LakePharma has a dedicated radioactive lab for conducting assays using H3, C14, P32, P33, S35, and I125. Whether you need a membrane-binding, scintillation... Show more »

Radiometric assays are the “gold standard” for many laboratories but establishing and maintaining these capabilities can be a burden for clients with limited resources. LakePharma has a dedicated radioactive lab for conducting assays using H3, C14, P32, P33, S35, and I125. Whether you need a membrane-binding, scintillation proximity (SPA), enzymatic, or other assay format involving a radioisotope, we have the expertise and facilities to assist you.

LakePharma’s Core Radiometric Assays include but are not limited to:

Radioligand Binding Assay
Radiometric Filter Binding Assay
SPA Based Assay
Autoradiography
Specific Activity Assay
Radioligand binding assays are a commonly used biochemical method of determining the binding characteristics of drug-like molecules which interact with transmembrane proteins. Common parameters include Kd and Bmax of radiolabeled molecules, and the IC50 and Ki of non-labeled competitor molecules.

SCOPE OF WORK:
1. The gene of interest is synthesized or amplified by PCR from a template provided by the client and cloned into our proprietary high expression Fc fusion vectors.
2. Pilot small-scale (0.03 liter) transient transfection into 293 cells is performed followed by protein A purification to assess expression levels and protein yield. Transient tranfection into CHO cells is available on request.
3. Expression is subsequently scaled up to 0.1 - 10 liters to produce starting material at the milligram to gram scale.
4. Purification is performed on a protein A column. Additional purification steps can be performed if required for additional cost.

A gene of interest or any desired DNA sequence can be synthesized; for DNA encoding proteins, codon optimization can be performed that can help improve expression in a variety of expression systems (bacterial, yeast, insect, mammalian). Restriction enzyme sites, promoters, and epitope tags can be incorporated anywhere for optimal... Show more »

A gene of interest or any desired DNA sequence can be synthesized; for DNA encoding proteins, codon optimization can be performed that can help improve expression in a variety of expression systems (bacterial, yeast, insect, mammalian). Restriction enzyme sites, promoters, and epitope tags can be incorporated anywhere for optimal utility. Sequence confirmation of the synthesized DNA is included as part of the service.

Gene Synthesis:
- Genes up to 500 bp = $200 total
- 6 day turnaround

-Genes 501 to 1000 bp = $0.50 per base
- 8 day turnaround

-Genes 1001 to 1500 bp = $0.60 per base
- 10 day turnaround

-Genes >1500 bp = Request Pricing
- Turnaround TBD

*NOTE: Complexity of sequence (GC-rich or repeated motifs) may result in additional costs. DNA sequencing and analysis is charged separately.

MATERIAL REQUIREMENT:
- DNA sequence to be synthesized as-is (if no optimization is performed)
- Protein sequence to be synthesized. Please note any flanking DNA sequences to be included, species for codon optimization, and whether specific restriction enzyme sites should be avoided.

DELIVERABLES: Plasmid DNA from a miniprep containing the sequence-verified synthesized DNA in the standard cloning plasmid, or an alternate plasmid specified and provided by the client. Note: Subcloning into a non-standard vector may result in additional costs.

SCOPE OF WORK:
Ligation cloning:
1. The gene of interest (provided in a plasmid or as a PCR product flanked by appropriate restriction enzyme sites) is digested with restriction enzymes and subcloned into the vector of interest containing compatible sites.
2. Plasmids containing insert are confirmed by restriction digest. The inserts can be sequenced for additional cost.

PCR cloning:
1. The gene of interest is amplified with primers containing appropriate restriction enzyme sites.
2. The PCR product is digested with restriction enzymes and subcloned into the plasmid of interest containing compatible sites.
3. Plasmids containing insert are confirmed by sequencing (for additional cost, see note below)
-Please note that DNA sequencing of the cloned PCR product is not included as a standard part of this service but is provided by LakePharma at an additional cost (see DNA sequencing and analysis link below). Sequencing the cloned PCR product is necessary to confirm that no mutations were introduced during PCR amplification.

Site-directed mutagenesis:
1. Primers are designed and used to perform the mutagenesis reactions.
2. Sequencing of the portion of the gene that was mutated is included to confirm that the desired change was incorporated.
-Please note that there will be an additional charge to sequence the remainder of the insert to ensure that no additional mutations were introduced during the mutagenesis reaction.

Gateway cloning:
1. A gateway LR recombination reaction is performed between the attL entry clone and the Gateway destination vector. (If a gateway entry clone is needed, this can be generated as a separate PCR cloning project.)
2. Clones with insert are verified by restriction enzyme digest or sequencing.

Custom cloning:
1. Additional sequences such as a peptide tag or protease cleavage site can be added using PCR.
2. Correct clones are identified by sequence analysis.

LakePharma has extensive experience with constructing and producing antibodies in our mammalian platforms. Antibody variable region genes (VH and VL) from a given antibody can be reformatted to encode a different constant region and other antibody-like molecules including Fab, scFv, and bispecific molecules. Multiple species and... Show more »

LakePharma has extensive experience with constructing and producing antibodies in our mammalian platforms. Antibody variable region genes (VH and VL) from a given antibody can be reformatted to encode a different constant region and other antibody-like molecules including Fab, scFv, and bispecific molecules. Multiple species and isotypes are available, with the option of synthesizing the constant region of choice. These expression constructs can be expressed in HEK293 and CHO cells followed by protein purification, and rapidly produced at scales ranging from milligram to gram quantities. We also offer high throughput 96-block antibody productions to fit your antibody screening needs.

SCOPE OF WORK:
1. Antibody genes are synthesized or amplified by PCR from a template provided by the client and cloned into our proprietary high expression mammalian vectors.
2. Pilot small-scale (0.03 liter) transient transfection into HEK293 is performed followed by protein A purification to assess expression levels and protein yield. Transient transfection into CHO is available on request.
3. Expression is subsequently scaled up to 0.25 - 20 liters to produce starting material at the milligram to gram scale.
4. Purification is performed on a protein A column. Additional purification steps can be performed if required for additional cost.

LakePharma offers custom services for CHO clonal stable cell line generation for a recombinant antibody or secreted protein. Proprietary stable cell line vectors are used as well as a suspension CHO cell line. Both a basic and a premium service are offered for clonal stable cell lines, depending on the client’s needs.

LakePharma offers custom services for CHO clonal stable cell line generation for a recombinant antibody or secreted protein. Proprietary stable cell line vectors are used as well as a suspension CHO cell line. Both a basic and a premium service are offered for clonal stable cell lines, depending on the client’s needs.

Clonal Cell Line Development (Basic) service includes:
1. Gene is subcloned into LakePharma’s proprietary mammalian expression vector system that contains multiple selectable markers.
2. CHO suspension cells are electroporated with expression constructs and placed under appropriate selection.
3. Clone selection up to 24 clones
4. Detailed characterization of the top clone during a 0.1 L production run (including viability, cell growth, and titer measurements)
5. Banking of 15 vials of the stable clone (10 million cells/vial)
6. Free backup vial storage for 6 months

Clonal Cell Line Development (Premium) service includes:
1. Gene is subcloned into LakePharma’s proprietary mammalian expression vector system that contains multiple selectable markers.
2. CHO suspension cells are electroporated with expression constructs and placed under appropriate selection.
3. Clone selection of up to 24 clones, including banking of top 3 clones for backup storage
4. Sub-Clone top 3 clones for a second round of selection including screening of up to 72 clones
5. Detailed characterization of the top 3 performing clones from the second round of selection during a 0.1 L production run of each (including viability, cell growth, and titer measurements)
6. Banking of 15 vials of the top performing clone (15 million cell/vial)
7. Detailed characterization of the final banked clone during a 1 L production run starting from thaw of a frozen vial (including viability, cell growth, and titer measurements)
8. Free backup vial storage for 12 months

DELIVERABLES:
FOR POOL CELLS:
- 10 vials of the final pool (5 million cells per vial, shipped on dry ice)
- 5 additional vials for backup storage at LakePharma (free for up to 6 months)
- Purified protein from 0.1 L production run
- Study Report

PREMIUM:
- 10 vials of the highest performing clone (15 million cells per vial, shipped on dry ice)
- 5 additional vials for backup storage at LakePharma (free for up to 1 year)
- Purified protein from all production runs, including 1 L production run of final banked clone
- Study Report

Small to large scale-up production of previously developed stable cell lines expressing proteins or antibodies. Custom protein purification protocols are not included, but can be designed based on the client’s needs for an additional charge

Small to large scale-up production of previously developed stable cell lines expressing proteins or antibodies. Custom protein purification protocols are not included, but can be designed based on the client’s needs for an additional charge